Differential regulation of cytokine-induced MMP-1 and MMP-13 expression by p38 kinase inhibitors in human chondrosarcoma cells: potential role of Runx2 in mediating p38 effects
Differential regulation of cytokine-induced MMP-1 and MMP-13 expression by p38 kinase inhibitors in human chondrosarcoma cells: potential role of Runx2 in mediating p38 effects
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DOI：
10.1016/j.joca.2006.01.017
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发表时间：
2006-08-01
影响因子：
7
通讯作者：
Thirunavukkarasu, Kannan
中科院分区：
文献类型：
--
作者：
Pei, Yong;Harvey, Anita;Thirunavukkarasu, Kannan
目的：研究丝裂原激活蛋白（MAP）激酶途径使用途径选择性抑制剂差异调节人软骨肉瘤细胞中基质金属蛋白酶（MMP）-1和-13表达的能力。设计：用细胞因子（IL-1β和TNFα）处理人软骨肉瘤细胞系（SW1353和JJ012）和人关节软骨细胞（HAC），并观察其表达情况。分析了MMP-1和-13。使用 ELISA 和蛋白质印迹分析评估 MAP 激酶抑制剂对细胞因子诱导的 MMP-1 和 -13 表达的影响。使用promote r-reporter检测、ELISA和免疫沉淀分析分析了Runx2通路可能参与介导p38对MMP-1 3表达的影响。结果：IL-1β和TNFα强烈诱导SW1353细胞和HAC中MMP-1和-13的表达，而在JJ012细胞中仅TNFa被发现诱导这两种MMP的表达。由于金属蛋白酶内源性组织抑制剂 (TIMP) 的过量产生，细胞因子治疗并未导致 MMP 活性显着增加。 p38激酶抑制剂（SB203580和SB242235）治疗以剂量依赖性方式强烈抑制细胞因子诱导的MMP-13表达，而对MMP-1表达的抑制作用稍弱。相比之下，细胞外信号调节激酶 (ERK) 和 c-Jun N 末端激酶 (JNK) 途径的抑制剂不会抑制任一 MMP 的表达。 Runx2 的过表达强烈刺激 MMP-1 3 的转录激活，但对 MMP-1 表达没有影响。此外，IL-1β 诱导 Runx2 磷酸化，而这种作用被 p38 激酶抑制剂阻断。我们的数据表明，Runx2 可能是 p38 效应在 IL-1 beta 诱导的 MMP-1 3 表达差异调节中的关键下游介质。结论：这些研究证明 p38 激酶抑制剂对人软骨肉瘤细胞中细胞因子诱导的 MMP-1 和 -13 表达的差异抑制。我们的研究还表明 Runx2 至少部分参与介导 p38 对 MMP-13 表达的影响。 (C) 2006 年国际骨关节炎研究协会。由爱思唯尔有限公司出版。保留所有权利。
Objective: To investigate mitogen activated protein (MAP) kinase pathways for their ability to differentially regulate the expression of matrix metalloprotease (MMP)-1 and -13 in human chondrosarcoma cells using pathway-selective inhibitors.Design: Human chondrosarcoma cell lines (SW1353 and JJ012) and human articular chondrocytes (HACs) were treated with cytokines (IL-1 beta and TNF alpha) and the expression of MMP-1 and -13 was analyzed. The effects of MAP kinase inhibitors on cytokine-induced expression of MMP-1 and -13 were evaluated using ELISA and Western blot analyses. The possible involvement of the Runx2 pathway in mediating p38 effects on MMP-1 3 expression was analyzed using promote r-reporter assays, ELISA and immunoprecipitation analyses.Results: IL-1 beta and TNF alpha strongly induced the expression of MMP-1 and -13 in SW1353 cells and HACs, whereas only TNFa was found to induce the expression of these two MMPs in JJ012 cells. Cytokine treatment did not result in a significant increase in the activity of MMPs because of the excess production of endogenous tissue inhibitors of metalloproteases (TIMPs). Treatment with p38 kinase inhibitors (SB203580 and SB242235) strongly inhibited cytokine-induced MMP-13 expression in a dose-dependent fashion while having a somewhat weaker inhibitory effect on MMP-1 expression. In contrast, inhibitors of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) pathways did not inhibit the expression of either MMP. Overexpression of Runx2 robustly stimulated the transcriptional activation of MMP-1 3 but had no effect on MMP-1 expression. Furthermore, IL-1 beta induced the phosphorylation of Runx2, and this effect was blocked by a p38 kinase inhibitor. Our data suggest that Runx2 is likely to be a key downstream mediator of p38 effects in the differential regulation of IL-1 beta induced MMP-1 3 expression.Conclusions: These studies demonstrate the differential inhibition of cytokine-induced MMP-1 and -13 expression by p38 kinase inhibitors in human chondrosarcoma cells. Our studies also suggest the involvement of Runx2, at least in part, in mediating the effects of p38 on MMP-13 expression. (C) 2006 OsteoArthritis Research Society International. Published by Elsevier Ltd. All rights reserved.